ACTIVE LEARNING SESSION

Calibration of IR Spectrophotometer

Represented by-:
17)Rushikesh Dake
19)Snehal Darade
19)Sanjana Deshmukh
20)Prasad Dethe

Subject – Advanced Instrumentation Techniques

Under guidance-: Prof.H.P. Khade Sir
 Calibration
• It demonstration that a particular instrument or device produces results within
specified limits by comparison with those produced by a reference or traceable
standard over an appropriate range of measurements.
Why should be Calibrated?

These should be calibrated for the following ways.

 To ensure the Reliability of the instrument.

 To determine the accuracy of the instrument.
 To ensure the Reading are consistent with other instrument.
 IR spectroscopy
• IR spectroscopy is the spectroscopy that deals with the infrared region of the
electromagnetic spectrum, that is light with a longer wavelength and lower
frequency than visible light.
• Infrared spectroscopy (IR) measures the bond vibration frequencies in a molecule
and is used to determine the functional groups.
• The infrared region of the spectrum encompasses radiation with wave numbers
ranging from about 12,500 to 50 cm -1 (or) wavelengths from 0.8 to 200 p.
• Infrared region lies between visible and microwave region.
• Infrared spectroscopy is the analysis of infrared light interacting with a molecule.
• It is based on absorption spectroscopy.
 Calibration Of IR Spectrophotometer
 Ensure that the FT-IR System is placed on a clean and dust free platform.
 Ensure that the instrument has been connected to power supply.
 Ensure that the all communication cords between the units are connected property.
 Dry indicator checks: The check must be done before turning the instrument on.
There are two indicators - an orange lamp as Dry indicator and green lamp as Power
indicator on the front. Dry indicator light shows the humidity in the interferometer
of the FT-IR even if it is off.
 Always confirm that the Dry indicator is on.
 Silica gel check: This check must also be done before turning the instrument on.
 Power spectrum check: This is done by clicking [BKG Scan] icon to perform
background scan measurements.
 A power spectrum is displayed after scanning. Click the Data tab and then
set scan parameters such as; the number of scans and resolution.
 Enter file name and sample information as "polystyrene film" and save in a
specific folder as "Calibration of FT-IR".
 Insert in the polystyrene film provided in the sample chamber cassette of the
FT-IR and start measurements.
 A typical spectrum of polystyrene will appear .
 Several pharmacopeias propose two tests for the resolution of Fourier
transform infrared(FT-IR) spectrometers using films of polystyrene.
 Polystyrene films have long been used for checking the general performance of
infrared(IR) spectrometers and specific tests for wave number accuracy and
resolution based on such films are enshrined in various pharmacopoeias.
 Instrument manufacturers supply polystyrene films for routine checks of their
spectrometers.
 The films have a matte finish that reduces the amplitude of interference fringes
but also increases scattering.
 Wave number verification
Prepare polystyrene film with thickness 0.04 mm and
verify the wave number.

Observed Wavenumber Minimum Tolerance Maximum Tolerance

3058.5 3061.5
2848.0 2851.0
1941.4 1944.4
1600.2 1602.2
1582.0 1584.0
1153.5 1155.5
1027.3 1029.3
 Resolution performance

Difference between % T at Difference between % T at

2851 cm -1 and 2870 cm-1 2583 cm- 1 and 1589 cm-1
Limit NLT 18 % NLS 12%
Actual
 They vary significantly in thickness, scattering properties, and even
chemical composition. Spectra of a number of such films are seen.
 The differences are unimportant for wave number accuracy tests
because the tolerances in the tests, typically ±1 cm, are much
greater than the variations seen between different commercial
polystyrene films.
 If required for compliance, calibrated polystyrene wave number
standards are available from sources such as, the National Institute
of Standards and Technology.